| Project/Area Number |
12660099
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bioproduction chemistry/Bioorganic chemistry
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| Research Institution | Fukui Prefectural University |
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Principal Investigator |
HIBI Takao Fukui Prefectural University, Dept. of Bioscience, Lecturer, 生物資源学部, 講師 (00285181)
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| Co-Investigator(Kenkyū-buntansha) |
TANIGUCHI Hiroshi Kwansei Gakuin Univ. School of Science & Technology, Assistant Professor, 理学部, 助教授 (10252719)
ODA Junichi Fukui Prefectural University, Dept. of Bioscience, Professor, 生物資源学研究科, 教授 (50027041)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | uricacid / thermostability / T-fold motif / intersubmit interraction / X-ray crystallography / Structural Biology / Oxidase / Uricase / Thermostability / 尿酸 / X線結晶構造解析 / 酸化還元酵素 / ウリカーゼ / X線結晶解析 / 酸素ラジカル |
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| Research Abstract |
Crystal structure of urate oxidase from Bacillus Sp. (bUOX) has been determined at 2.2A resolution. X-Ray diffraction data for the crystals were collected using a Mar CCD camera and synchrotron radition on beam line BL41XU (Spring-8, Harima, Japan). The crystals belong to space group P2_12_12 with unitcell parameters a=133.5, b=144.5, c=78.8A.
The structure of UOX-8-azaxanthine complex has been solved by molecular replacement techniques. The refined structure of UOX from Asperigillus flavus (PDS code 1UOX; Colloc'h et al. (1997)) was used as our initial model, all the residues of which were replaced with alanines. Restrained refinement with Refmac5 (CCP4 Project No.4 (1999)) resulted in an R-value of 22% for the data (PDB code: 1J2G). Main chain structures of the loops at tetrametric interface are significantly different from those of A. flavus UOX.
Instability of the b UOX crystal quality in changing salt concentration prevents from preparing Xe derivatives. However, another salt effect to bUOX was found that its thermal stability increases with salt concentration. Interestingly, one of the loops at the tetrameric interface contains R298 and a sulfate ion seems to bridge the subunits related with a non-crystallographic two-fold sysmmetry.
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